论文部分内容阅读
研究活化T细胞核因子(NFAT)在环境内分泌干扰物双酚A(BPA)调控小鼠单核巨噬细胞白血病细胞(RAW264.7)分泌白细胞介素-10(IL-10)蛋白中的作用,为进一步研究BPA的免疫毒作用机制提供理论依据。将不同剂量的BPA(0.1、1、10、100及1000μmol/L)作用于RAW264.7细胞24 h、48 h,用CCK8试剂盒检测细胞活性。以1μg/ml脂多糖(LPS)为激活剂,以1、10、50、100μmol/L BPA作用细胞24 h,ELISA试剂盒检测细胞上清液中IL-10蛋白含量。10、100μmol/L BPA作用细胞4 h、12 h,用实时定量PCR方法测定IL-10、NFATc、NFATp基因表达。与对照组比较,1000μmol/L BPA无论作用24 h还是48 h均能明显抑制细胞活性(P<0.01);LPS显著促进巨噬细胞分泌IL-10蛋白,50和100μmol/L BPA作用细胞24 h,与LPS组比较,显著抑制IL-10蛋白的分泌(P<0.01);10μmol/L、100μmol/L BPA染毒4 h能明显促进NFATp mRNA表达(P<0.01);染毒12 h时,IL-10 mRNA水平降低,差异有统计学意义(P<0.05)。提示本实验条件下,NFATp在BPA调控巨噬细胞分泌IL-10蛋白中具有一定的作用。
To investigate the role of activated nuclear factor-kappaB (NFAT) in the secretion of interleukin-10 (IL-10) by the environmental endocrine disruptor BPA in mouse monocyte-macrophage leukemia cells (RAW264.7) Provide a theoretical basis for further study of the immune toxicity mechanism of BPA. Different doses of BPA (0.1, 1, 10, 100 and 1000 μmol / L) were applied to RAW264.7 cells for 24 h and 48 h, respectively. Cell viability was detected by CCK8 kit. The cells were treated with 1, 10, 50 and 100μmol / L BPA for 1 h at 1μg / ml lipopolysaccharide (LPS) as an activator, and IL-10 protein content in supernatant was detected by ELISA kit. After treated with 10,100μmol / L BPA for 4 h and 12 h, the expression of IL-10, NFATc and NFATp was detected by real-time PCR. Compared with the control group, 1000μmol / L BPA could significantly inhibit the cell viability at 24 h and 48 h (P <0.01). LPS significantly promoted the secretion of IL-10 protein by macrophages, and at 50 and 100 μmol / L BPA for 24 h (P <0.01). Compared with the LPS group, the secretion of IL-10 protein was significantly inhibited (P <0.01). The expression of NFATp mRNA was significantly increased after treated with 10μmol / L or 100μmol / L BPA for 4 h IL-10 mRNA levels decreased, the difference was statistically significant (P <0.05). It is suggested that NFATp may play a role in the secretion of IL-10 protein by BPA regulated macrophages in this experiment.